Mx3000p apparatus
The Mx3000P is a real-time PCR (qPCR) system designed for gene expression analysis and nucleic acid quantification. It features a compact design, intuitive software, and a range of sample block options to accommodate various sample volumes and formats. The Mx3000P provides precise temperature control, sensitive optical detection, and robust data analysis capabilities to support a variety of real-time PCR applications.
4 protocols using mx3000p apparatus
RNA Extraction and qRT-PCR Analysis
Quantitative Gene Expression Analysis
Extraction and Quantification of Total RNA
RNA quantity and purity were determined using a spectrophotometer (DeNovix Inc., Wilmington, DE, USA). Total RNA (1 μg) from each sample was reverse-transcribed using hexanucleotides. Real-time RT–PCR measurements were performed with an Mx3000P apparatus (Agilent Technologies, Santa Clara, CA, USA) with the corresponding SYBR Green kit. PCR primers were designed with the Primer Blast or Primer BD programs and obtained from Promega, Madisson, WI, USA. Gene expression was normalized to β-actin. All primers used are indicated in
Quantifying VEGF Expression in CRC Cells
Western blot analysis was carried out as described previously [42 (link)]. The primary antibodies were directed against HIF-1alpha (BD Bioscience # 610958), HIF-2alpha (Novus Biological # NB-100122) or beta-actin (Santa Cruz # sc-47778), while the corresponding secondary antibodies were purchased from Jackson ImmunoResearch.
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